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Effect of Dietary Zinc Deficiency on the Endogenous Phosphorylation and Dephosphorylation of Rat Erythrocyte Membrane^1,2,

Department of Nutritional Sciences, College of Biological Science ^* Department of Mathematics and Statistics, University of Guelph, Guelph, Ontario, Canada N1G 2W1

The effect of dietary zinc deficiency on patterns of phosphorylation and dephosphorylation of rat erythrocyte membrane proteins and erythrocyte filterability was examined. Weanling male Wistar rats were fed an egg white—based diet containing less than 1.1 mg zinc/kg diet ad libitum for 3 wk. Control rats were either pair-fed or ad libitum—fed the basal diet supplemented with 100 mg zinc/kg diet. Net phosphorylation and dephosphorylation of erythrocyte membrane proteins were carried out by an in vitro assay utilizing [-³²P]ATP. The membrane proteins were subsequently separated by sodium dodecyl sulfate—polyacrylamide gel electrophoresis, and the ³²P content of gel slices was counted by Cerenkov counting. Erythrocyte filterability was measured as the filtration time of suspensions of erythrocytes, both untreated and preincubated with diamide, under constant pressure. Erythrocyte ghosts from zinc-deficient rats demonstrated greater dephosphorylation of protein bands R1 plus R2 and R7 than pair-fed rats and greater net phosphorylation of band R2.2 than pair-fed or ad libitum—fed control rats (P < 0.05). Erythrocytes from ad libitum-fed control rats showed significantly longer filtration times than those from zinc-deficient or pair-fed control rats. In conclusion, dietary zinc deficiency alters in vitro patterns of erythrocyte membrane protein phosphorylation and dephosphorylation, whereas the depression in food intake associated with the zinc deficiency increases erythrocyte filterability.

KEY WORDS: • zinc deficiency • erythrocyte membrane • protein phosphorylation/dephosphorylation

¹ This research was supported by the Natural Sciences and Engineering Research Council of Canada and the Ontario Ministry of Agriculture and Food.

² From a thesis submitted by Phyllis Paterson in partial fulfillment of the requirements for the degree of Doctor of Philosophy in Nutritional Sciences at the University of Guelph.

Manuscript received 10 April 1987. Revision accepted 12 August 1987.